Vascular disease is a leading cause of death and disability in the western world. Diagnosis and staging of atherosclerosis is a challenge, especially with regards to the identification of plaque vulnerability. We are developing imaging methods based upon MRI and intravascular microcoils. In order to rigorously validate our MRI imaging methods and algorithms, we have developed a new cryo-imaging system that allows one to alternately section and image the block face of tissue. We obtain 3D pathology of vessel segments excised from cadaver and we characterize the tissues of atheroma using episcopic autofluorescence and bright field microscopy images. After embedding the vessel, the block is frozen, and block face microscopic images are taken every 200μm with an image resolution of 30μm×30μm. The series of images is then corrected for uneven illumination, serially registered to one another, and the 3D vessel segment is reconstructed. Some sections are recovered and processed with histological staining for validation. Seven tissue types can be readily identified from the cryo-images: necrotic core, calcification, lipid pool, media, adventitia, fibrosis, thrombus, and normal intima. Since the whole vessel segment is available, we could register 3D data to images from MR, or other modalities, for validation. In addition, visualization tools such as multi-planar reformatting 3D rendering can be used to study 3D plaque morphology, in microscopic detail.